Work

Fully automated sample to result SIMPLE RPA microfluidic chip towards in ovo sexing application

Year: 2024

Type: article

Abstract: Several European countries have implemented new legislations to eliminate the killing of day-old male chicks, pushing the laying hen industry to find animal-friendly solutions. Although embryo sexing during incubation (in ovo sexing) is highly promising, no current solution meets the industry requirements of handling all egg colors with >98 % sexing accuracy, low cost and minimal embryo disturbance, while allowing sexing before day 13 of incubation and processing > 20 000 eggs/hour. Recombinase polymerase amplification (RPA) presents a promising alternative to PCR as a sensitive isothermal technique to be integrated into a microfluidic platform. In this work we developed a fully autonomous microfluidic cartridge (SIMPLE-RPA chip) for chick female-specific synthetic HINTW gene detection in 30 minutes at 37.7 °C inside an egg incubator. We first optimized off-chip RPA, allowing for highly sensitive DNA detection (1.6 x 10-5 ng/µL), matching expected concentrations in allantoic fluid samples. Then, the SIMPLE-RPA chip was developed to accommodate and automate the RPA bioassay on-chip, requiring only sample introduction and finger press activation to reduce user errors and contamination risks. Importantly, the SIMPLE-RPA chip maintained the LOD achieved off-chip. The presented SIMPLE-RPA chip is inexpensive, has a small footprint, is compatible with upscaling, and can be easily transferred to other point-of-care applications.

Source: npj Biosensing

DOI: 10.1038/s44328-024-00019-6